白芍总苷对角质形成细胞增殖及血管内皮生长因子和白介素-23表达的影响

目的 研究白芍总苷（TGP）对角质形成细胞增殖和分泌血管内皮生长因子（VEGF）和白介素（IL）-23的影响,探讨可能涉及的信号转导通路。方法 不同浓度TGP作用于体外培养的HaCaT细胞株,噻唑蓝（MTT）法观察TGP对HaCaT细胞增殖活性的影响。将HaCaT细胞分为三组,即对照组不加任何刺激因素,TGP组分别加入6种不同浓度的TGP,SB203580组在加入10 mol/L SB203580预处理2 h后加入125 mg/L TGP。实时定量PCR（RT-PCR）方法和ELISA方法检测TGP对HaCaT细胞VEGF和IL-23表达的影响;免疫印迹技术观察TGP作用于HaCaT细胞后p38的磷酸化及SB203580对p38磷酸化的影响。结果 TGP在低浓度（0.5、2.5 mg/L）时对HaCaT细胞增殖有促进作用,浓度≥12.5 mg/L时反而对细胞的增殖有抑制作用,至125 mg/L时抑制作用最强。TGP在低浓度（0.5、2.5 mg/L）时对HaCaT细胞VEGF和IL-23 mRNA和蛋白的表达有促进作用,12.5 ～ 125 mg/L时内可抑制HaCaT细胞VEGF mRNA和蛋白的表达,62.5 ～ 125 mg/L时可抑制IL-23 mRNA和蛋白的表达。TGP可时间依赖性地诱导HaCaT细胞p38的磷酸化,磷酸化p38 蛋白于125 mg/L TGP作用5 min后达到高峰,表达水平为0.3314 ± 0.0245,10 min后减弱至0.2173 ± 0.0189,但均高于对照组水平;30 min后表达水平降为0.1664 ± 0.0201;SB203580可减弱其作用,SB203580预处理组磷酸化p-p38 表达水平为0.1529 ± 0.0147。结论 TGP可抑制HaCaT细胞的增殖及VEGF和IL-23 mRNA和蛋白的表达,p38MAPK信号途径可能介导其抑制作用。     

Effects of tacrolimus on the expression of nuclear factor-κB and glucocorticoid receptor by HaCaT cells in vitro

Objective To investigate the effect of tacrolimus on the expression of nuclear factor-κB (NF-κB) in HaCaT cells stimulated by tumor necrosis factor-α(TNF-α),and on the expression of glucocorticoid receptor (GR)α and β in untreated HaCaT cells in vitro.Methods Cultured Ha CaT cells were treated with TNF-α(10μg/L) only,combination of TNF-α(10μg/L) and various concentrations (10-8mol/L, 10-7mol/L,10-6moL/L) of tacrolimus or tacrolimus of different concentrations only.After additional 12-,24-, 36- or 48-hour cnlture, Westem blot and immunofluorescenee-confocal laser scanning microscopy were used to detect the expressions of NF-κB,GRα and GRβ in HaCaT cells.Those untreated HaCaT cells served as the control.Results The relative protein expression level of NF-κB was increased in HaCaT cells after treatment with TNF-α for 24 and 48 hours zompared with untreated ceils (0.73±0.0316 and 0.8925±0.0171 vs 0.4988±0.03506,both P<0.05);however,the increase in NF-κB expression was inhibited by the combination treatment with tacrolimus,and the relative expression level of NF-κB protein was 0.6825±0.0263.0.6200±0.0163 and 0.5575±0.0299 in HaCaT cells treated with TNF-α plus tacrolimus of 10-8mol/L 10-7mol/L and 10-6mol/L,respectively;the difference was significant etween TNF-α-treated cells and those dealt with the combination of NF-α and tacrolimus of 10-7 or 10-6 mol/L (both P<0.05).No significant difference was observed in the expression of NF-κB by HaCaT cells between different time oints treated with tacrolimus of 10-8,10-7 or 10-6 mol/L.Also,there was no zignificant difference in the expression of GRα or GRβ between untreated HaCaT cells and those treated with tacrolimus of 10-8, 10-7 or 10-6 mol/L at any time point.Conclusions Tacrolimus ould inhibit the expression of NF-κB by TNF-α-stimulated HaCaT cells,but does not affect the expression of GRα or GRβ,in untreated HaCaT cells.     

Effects of tacrolimus on the expression of nuclear factor-κB and glucocorticoid receptor by HaCaT cells in vitro

Objective To investigate the effect of tacrolimus on the expression of nuclear factor-κB (NF-κB) in HaCaT cells stimulated by tumor necrosis factor-α(TNF-α),and on the expression of glucocorticoid receptor (GR)α and β in untreated HaCaT cells in vitro.Methods Cultured Ha CaT cells were treated with TNF-α(10μg/L) only,combination of TNF-α(10μg/L) and various concentrations (10-8mol/L, 10-7mol/L,10-6moL/L) of tacrolimus or tacrolimus of different concentrations only.After additional 12-,24-, 36- or 48-hour cnlture, Westem blot and immunofluorescenee-confocal laser scanning microscopy were used to detect the expressions of NF-κB,GRα and GRβ in HaCaT cells.Those untreated HaCaT cells served as the control.Results The relative protein expression level of NF-κB was increased in HaCaT cells after treatment with TNF-α for 24 and 48 hours zompared with untreated ceils (0.73±0.0316 and 0.8925±0.0171 vs 0.4988±0.03506,both P<0.05);however,the increase in NF-κB expression was inhibited by the combination treatment with tacrolimus,and the relative expression level of NF-κB protein was 0.6825±0.0263.0.6200±0.0163 and 0.5575±0.0299 in HaCaT cells treated with TNF-α plus tacrolimus of 10-8mol/L 10-7mol/L and 10-6mol/L,respectively;the difference was significant etween TNF-α-treated cells and those dealt with the combination of NF-α and tacrolimus of 10-7 or 10-6 mol/L (both P<0.05).No significant difference was observed in the expression of NF-κB by HaCaT cells between different time oints treated with tacrolimus of 10-8,10-7 or 10-6 mol/L.Also,there was no zignificant difference in the expression of GRα or GRβ between untreated HaCaT cells and those treated with tacrolimus of 10-8, 10-7 or 10-6 mol/L at any time point.Conclusions Tacrolimus ould inhibit the expression of NF-κB by TNF-α-stimulated HaCaT cells,but does not affect the expression of GRα or GRβ,in untreated HaCaT cells.     

CpG ODN刺激的寻常性银屑病中性粒细胞培养上清液对HaCaT细胞增殖的影响

Objective To investigate the role of nentrophils in the pathogenesis of psoriasis vulgaris. Methods Neutrophils were isolated from venous blood samples of 25 patients with psoriasis vulgaris (including 13 cases of active psoriasis and 12 cases of inactive psoriasis) as well as 25 normal human con-trols, and cultured. Then, these neutrophils were grouped and treated with lipopolysaccharide (LPS, 100 g/L),CpG-A (50 mg/L), CpG-B (50 mg/L), and RPMI 1640 culture medium, respectively, for 24 hours followed by the collection of culture supematants. Human keratinocytes (HaCaT) were cultured in the presence of su-pematants of treated or untreated nentrophils for 72 hours followed by the detection of cell proliferation with MTT assay. To determine the role of proinflammatory factors, SOD/CAT and monoclonal antibody to IL-8 and TNF-alpha of 400 u/mL were used to pretreat HaCaT cells 1 hour prior to the stimulation with super-natants of neutrophils. Results Compared with culture medium, the supematant of unstimulated neutrophils from normal controls or patients with inactive psoriasis had no significant effect on the proliferation of HaCaT cells (P ＞ 0.05), but that from patients with active psoriasis markedly promoted the proliferation of HaCaT cells (t = 2.41, P < 0.05). ARe, stimulation by LPS, CpG-A and CpG-B, the supematant of active patient-derived neutrophils significantly promoted the proliferation of HaCaT cells compared with that of normal control-derived nentrophils (t = 3.11, 2.89, 2.29, respectively, all P < 0.05). In comparison with tmstimulated neutrophils, the supematant from LPS- and CpG-A stimulated nentrophiles significantly accelerated the pro-liferation of HaCaT cells. Furthermore, the proliferation of HaCaT cells induced by the supematants of LPS-,CpG-A-, CpG-B-stimulated neutrophils from psoriatic patients was statistically suppressed by the pretreat-ment with the monoclonal antibody to IL-8, TNF-alpha and SOD/CAT (all P < 0.05). Conclusions In patients with psoriasis vulgaris, there is an abnormal secretion of IL-8, TNF-alpha and superoxide by neutrophils in peripheral blood, and these proinflammatory factors could promote the proliferation of HaCaT cells.     

CpG ODN刺激的寻常性银屑病中性粒细胞培养上清液对HaCaT细胞增殖的影响

Objective To investigate the role of nentrophils in the pathogenesis of psoriasis vulgaris. Methods Neutrophils were isolated from venous blood samples of 25 patients with psoriasis vulgaris (including 13 cases of active psoriasis and 12 cases of inactive psoriasis) as well as 25 normal human con-trols, and cultured. Then, these neutrophils were grouped and treated with lipopolysaccharide (LPS, 100 g/L),CpG-A (50 mg/L), CpG-B (50 mg/L), and RPMI 1640 culture medium, respectively, for 24 hours followed by the collection of culture supematants. Human keratinocytes (HaCaT) were cultured in the presence of su-pematants of treated or untreated nentrophils for 72 hours followed by the detection of cell proliferation with MTT assay. To determine the role of proinflammatory factors, SOD/CAT and monoclonal antibody to IL-8 and TNF-alpha of 400 u/mL were used to pretreat HaCaT cells 1 hour prior to the stimulation with super-natants of neutrophils. Results Compared with culture medium, the supematant of unstimulated neutrophils from normal controls or patients with inactive psoriasis had no significant effect on the proliferation of HaCaT cells (P ＞ 0.05), but that from patients with active psoriasis markedly promoted the proliferation of HaCaT cells (t = 2.41, P < 0.05). ARe, stimulation by LPS, CpG-A and CpG-B, the supematant of active patient-derived neutrophils significantly promoted the proliferation of HaCaT cells compared with that of normal control-derived nentrophils (t = 3.11, 2.89, 2.29, respectively, all P < 0.05). In comparison with tmstimulated neutrophils, the supematant from LPS- and CpG-A stimulated nentrophiles significantly accelerated the pro-liferation of HaCaT cells. Furthermore, the proliferation of HaCaT cells induced by the supematants of LPS-,CpG-A-, CpG-B-stimulated neutrophils from psoriatic patients was statistically suppressed by the pretreat-ment with the monoclonal antibody to IL-8, TNF-alpha and SOD/CAT (all P < 0.05). Conclusions In patients with psoriasis vulgaris, there is an abnormal secretion of IL-8, TNF-alpha and superoxide by neutrophils in peripheral blood, and these proinflammatory factors could promote the proliferation of HaCaT cells.     

CpG ODN刺激的寻常性银屑病中性粒细胞培养上清液对HaCaT细胞增殖的影响

Objective To investigate the role of nentrophils in the pathogenesis of psoriasis vulgaris. Methods Neutrophils were isolated from venous blood samples of 25 patients with psoriasis vulgaris (including 13 cases of active psoriasis and 12 cases of inactive psoriasis) as well as 25 normal human con-trols, and cultured. Then, these neutrophils were grouped and treated with lipopolysaccharide (LPS, 100 g/L),CpG-A (50 mg/L), CpG-B (50 mg/L), and RPMI 1640 culture medium, respectively, for 24 hours followed by the collection of culture supematants. Human keratinocytes (HaCaT) were cultured in the presence of su-pematants of treated or untreated nentrophils for 72 hours followed by the detection of cell proliferation with MTT assay. To determine the role of proinflammatory factors, SOD/CAT and monoclonal antibody to IL-8 and TNF-alpha of 400 u/mL were used to pretreat HaCaT cells 1 hour prior to the stimulation with super-natants of neutrophils. Results Compared with culture medium, the supematant of unstimulated neutrophils from normal controls or patients with inactive psoriasis had no significant effect on the proliferation of HaCaT cells (P ＞ 0.05), but that from patients with active psoriasis markedly promoted the proliferation of HaCaT cells (t = 2.41, P < 0.05). ARe, stimulation by LPS, CpG-A and CpG-B, the supematant of active patient-derived neutrophils significantly promoted the proliferation of HaCaT cells compared with that of normal control-derived nentrophils (t = 3.11, 2.89, 2.29, respectively, all P < 0.05). In comparison with tmstimulated neutrophils, the supematant from LPS- and CpG-A stimulated nentrophiles significantly accelerated the pro-liferation of HaCaT cells. Furthermore, the proliferation of HaCaT cells induced by the supematants of LPS-,CpG-A-, CpG-B-stimulated neutrophils from psoriatic patients was statistically suppressed by the pretreat-ment with the monoclonal antibody to IL-8, TNF-alpha and SOD/CAT (all P < 0.05). Conclusions In patients with psoriasis vulgaris, there is an abnormal secretion of IL-8, TNF-alpha and superoxide by neutrophils in peripheral blood, and these proinflammatory factors could promote the proliferation of HaCaT cells.     

紫外线致皮肤光损伤皮肤癌的机制

波长范围在280nm～320nm的中波紫外线（UVB）在光损伤皮肤癌的发展中占有重要的作用。它主要被表皮吸收,尤其表现为影响机体的免疫系统、凋亡分子等,从而诱发皮肤癌。     

Cr(III) and Cr(VI) in leather and elicitation of eczema

The aim of the present study was to investigate the relation between the content of Cr(VI) and soluble Cr(III) in leather and the ability of the leather to elicit eczema in chromium allergic patients. An array of chromium-tanned leather samples was analysed for the content of total Cr(VI) and soluble Cr(III) using the DIN 53314 and the DS/EN 420 methods. Subsequently, a group of 15 patients with a history of foot eczema and leather exposure was exposed to a selection of 14 chromium- and 1 vegetable-tanned leather sample on the upper back for 48 hr. In addition, one leather sample was used for a prolonged 14-day exposure study. In total, 4 of the 15 patients reacted to at least one leather sample, and 5 of the 14 leather samples elicited a reaction in at least 1 patient. The prolonged exposure study demonstrated that an extended exposure period may reveal allergenic potential of a leather sample not otherwise identified using an ordinary 48-hr exposure period. No relation was observed between the measured content of Cr(VI) and soluble Cr(III) in the leather and the elicitation of eczema. Thus, in order to evaluate the quality of chromium-tanned leather in relation to preventing allergic skin reactions, other more clinical relevant methods reflecting the actual bioavailable Cr(III) and Cr(VI) fractions should be developed.     

Pseudo-angiosarcomatosis (kaposi) and arterio-venous fistulae (author's transl)

In a few cases, arterio-venous fistulae of the lower limbs lead to cutaneous symptoms of the toes, feet or legs exactly similar to Kaposi's disease; the fairly precocious occurrence, the unilateral localisation, the existence of clinical and radiological arterial symptoms and usually, but not always, particular histological features allow this discrimination. Seen in both sex, male prevailing, between 12 and 50, it can simulate Kaposi's disease even on histological grounds; the diagnosis may, then, be difficult inasmuch as vascular symptoms may clinically lack. The development may be very different from one case to another so that there is no standard treatment between "wait and see" and more or less extensive amputation.     

Surfactant protein D (SP-D) and systemic scleroderma (SSc)

We measured serum levels of SP-D in collagen diseases (110 cases) such as systemic scleroderma (SSc), scleroderma spectrum disorders (SSD), systemic lupus erythematodes (SLE), Sjogren syndrome (Sjs), dermatomyositis (DM), rheumatoid arthritis (RA), and dermatitis (DE) (109 cases) as a control. Additionally, we performad a correlation analysis to determine how these levels were related to pulmonary fibrosis and function test (vital capacity, %DLco). The serum levels of SP-D increased in SSc patients with Barnett type III more than in SSc patients with Barnett type I or II, while they increased slightly in SSD (incomplete type of SSc) patients. The differences in these figures were statistically significant between the SSc (SSc & SSD) and non-SSc (SLE, DM, Sjs & RA) groups (p<0.005). The serum levels of SP-D in SSc patients with anti-topoisomerase I antibodies were statistically higher than those in SSc patients with other types of anti-nuclear antibodies. There was a statistically significant correlation between the severity of pulmonary fibrosis and the serum levels of SP-D, and a statistically negative correlation between SP-D levels and vital capacity or %DLco, but there was no proportional correlation with the forced expiatory volume (FEV1.0%). There was no statistical relationship between pre- and post-therapy with photopheresis; however, there was a statistical correlation between the serum levels of SPD and KL-6. In the group of collagen diseases, plasma levels of SP-D were higher than serum levels of SP-D. Patients with SSc possess higher levels of SP-D than do those with other collagen diseases and dermatitis, which may correspond to the severity of pulmonary fibrosis.